| In the review of this thesis, the basic principles, characteristics and the applications of chemiluminescence are briefly introduced. The flow injection chemiluminescence (FI-CL) is becoming more and more popular owing to its simple instrumentation, high sensitivity, wide dynamic range, reproducibility, simplicity and rapidly. It is well known that pharmaceutical analysis has taken great important role in clinical, pharmacokinetic research. It is not only of great theoretical meaning but also of application value that chemiluminescence is applied in the pharmaceutical analysis. New flow injection chemiluminescene methpds coupled with other techniques such as immobilized enzyme were built for the determination of Triglyceride and cholesterol. This thesis chose hemoglobin as typical enzyme of peroxidase and mimetic perosidase. The research section includes four subsections as following:1. A post chemiluminescence phenomenon was observed when Betahistine Hydrochloride solution was injected into a solution with alkaline luminal and potassium periodate which had been CL reaction. A possible reaction mechanism was proposed. Using the Post chemiluminescence reaction, a new method for the determination of Betahistine Hydrochloride was established by flow injection. The linear response range of this method was 2×10-7-4×10-4 mol/L with a linear correlation coefficient of 0. 9975. The limit of detection was 1.0×10-7 mol/L. The relative standard deviation for 4.0×10-5 mol/L solution was 2.9 % (n = 11).2. Based on the reduction of hexavalent chromium by itopide hydrochloride to trivalent chromium, which in turn catalyzed the chemiluminescence of luminal-H2O2, a rapid, sensitive and new method for the determination of itopide hydrochloride was established. The linear range of 1.0×10-11-1.0×10-9 mol/L for itopide hydrochloride was found with a detection limit of 2.0×10-12 mol/L. The relative standard deviation (n = 11) for 1.65×10-10mol/L itopide hydrochloride was 2.5%. This method was successfully applied to the determination of itopide hydrochloride in the tablets.3. With the catalyst of hemoglobin, the chemiluminescence was yielded by the luminol and H2O2 released from the reaction between TG and its relative enzymes. A line relationship between the CL intensity and hydrogen peroxide was observed in certain of range. Because of these, a new indirect method detecting TG of human serum was established. The linear response range of this method was 2.26×10 -4-2.26×10-6 mol/L with a linear correlation coefficient of 0.9955. The limit of detection was 7.5×10-7mol/L. The compared result between this method and traditional enzyme method has not significant difference by calculated.4. Cholesterol oxydase and cholesterol esterase were immobilized in the course of gelation of the sol of tetraethyl orthosilicate, and immobilized columns were easily prepared with silica gel using this system. When the sample flew across the column, hydrogen peroxide was released. With the catalyst of hemoglobin, the chemiluminescence was yielded by the luminol and H2O2. A line relationship between the CL intensity and cholesterol concentration was observed in the certain of range. Because of these, a new indirect method detecting cholesterol (including free cholesterol and total cholesterol) of human serum was established. The linear response range of this method to total cholesterol was 2.02×10-4-1.01×10-6 mol/L with a linear correlation coefficient of 0.9975. The limit of detection was 3.5×10-7 mol/L. The linear response range of this method to free cholesterol was 2.18×10 -5-5.0×10-8 mol/L with a linear correlation coefficient of 0.9991. The limit of detection was 5.0×10-9 mol/L. The results from this method and traditional enzyme methods have not significant difference by calculated. |
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